Hostname: page-component-7c8c6479df-hgkh8 Total loading time: 0 Render date: 2024-03-19T06:20:57.131Z Has data issue: false hasContentIssue false

Isolation and properties of an RNA fraction present in Brucella culture supernatants

Published online by Cambridge University Press:  15 May 2009

M. J. Corbel
Affiliation:
Ministry of Agriculture, Fisheries and Food, Central Veterinary Laboratory, New Haw, Weybridge, Surrey
R. A. Brewer
Affiliation:
Ministry of Agriculture, Fisheries and Food, Central Veterinary Laboratory, New Haw, Weybridge, Surrey
Rights & Permissions [Opens in a new window]

Summary

Core share and HTML view are not available for this content. However, as you have access to this content, a full PDF is available via the ‘Save PDF’ action button.

The supernatant fluids of batch and continuous cultures of Brucella strains contained up to 100 mg/i of soluble RNA which could be recovered by precipitation with lysozyme. This RNA fraction had many of the properties of ribosomal RNA and was single-stranded, sensitive to ribonuclease, with an approximate sedimentation constant of 5S, a molecular weight of about 35000 daltons and an adenine; guanine; cytosine; uracil content of 17·5 26·5 33; 23 mol% respectively. RNA fractions from lysozyme precipitates evoked high titres of Brucella agglutinins on injection into rabbits and induced acute inflammatory responses in guinea-pig skin. Highly purified RNA fractions prepared by phenol extraction of lysozyme precipitates did not evoke antibodies to Brucella abortus.

Type
Research Article
Copyright
Copyright © Cambridge University Press 1980

References

REFERENCES

Alton, G. C. & Jones, L. M. (1967). Laboratory techniques in brucellosis. World Health Organization Monograph Series, No. 55.Google Scholar
Bendich, A. (1957). Methods for characterization of nucleic acids by base composition. In Methods in Enzymology, vol. 3 (ed. Colowick, S. P. and Kaplan, N. O.), pp.715–23. New York: Academic Press.Google Scholar
Boyce, K. J. & Edgar, A. W. (1966). Production of freeze.dried Brucella abortus strain 19 vaccine using cells produced by continuous culture. Journal of Applied Bacteriology 29, 401–8.CrossRefGoogle Scholar
Burnet, E. (1922). Sur un nouveau procédé de diagnostic dela fièvre méditerranéenne. Compte rendu de l'Academie des Sciences, Paris 174, 421–3.Google Scholar
Corbel, M. J. (1973). Evaluation of an immunodiffusion test for the detection of antibodies to Brucella abortus in bovine serum. Journal of Medical Microbiology 6, 6776.Google Scholar
Corbel, M. J. (1976 a). The immunological properties of brucella ribosomal preparations. Developments in Biological Standardization 31, 115–32.Google Scholar
Corbel, M. J. (1976 b). The immunogenic activity of ribosomal fractions derived from Brucella abortus. Journal of Hygiene 76, 6574.CrossRefGoogle ScholarPubMed
Corbel, M. J. (1977). Isolation and partial characterization of a phage receptor from Brucella neotomae 5K33. Annali Sclavo 19, 131–42.Google Scholar
Davis, B. J. (1964). Disc electrophoresis. II. Method and application to human serum proteins. Annals of the New York Academy of Sciences 121, 404–27.Google Scholar
Dische, Z. (1947). A new specific color reaction of hexuroruc acids. Journal of Biological Chemistry 167, 189–98.CrossRefGoogle ScholarPubMed
Dubois, M., Gilles, K. A., Hamilton, J. K., Rebers, P. A. & Smith, F. (1956). Colorimetric method for determination of sugars and related substances. Analytical Chemistry 28, 350–6.CrossRefGoogle Scholar
Ellwood, D. C., Keppie, J. & Smith, H. (1967). The chemical basis on the virulence of Brucella abortus. VIII. The identity of purified immunogenic material from culture filtrate and from the cell wall of Brucella abortus grown in vitro. British Journal of Experimental Pathology 48, 2839.Google Scholar
Fraenkel-Conrat, H. (1954). Reaction of nucleic acid with formaldehyde. Biochimica et Biophysica Acta 15, 307–9.CrossRefGoogle ScholarPubMed
Freeman, B. A., McGhee, J. R. & Baughn, R. E. (1970). Some physical, chemical and taxonomic features of the soluble antigens of the Brucellae. Journal of Infectious Diseases 121, 522–7.CrossRefGoogle ScholarPubMed
Giles, K. W. & Myers, A. (1965). An improved diphenylamine method for the estimation of deoxyribonucleic acid. Nature, London 206, 93.CrossRefGoogle Scholar
Janda, J. & Work, E. (1971). A colorimetric estimation of lipopolysaccharides. Federation of European Biochemical Societies Letters 16, 343–5.CrossRefGoogle ScholarPubMed
Jones, L. M., Diaz, R. & Taylor, A. G. (1973). Characterization of allergens prepared from smooth and rough strains of Brucella melitensis. British Journal of Experimental Pathology 54, 492508.Google ScholarPubMed
Kabat, E. A. & Mayer, M. M. (1961). In Kabat and Mayer‘s Experimental Immunochemistry 2nd ed, ch. 22: Carbohydrate estimation, pp. 526–37. Springfield, Illinois: Charles C. Thomas.Google Scholar
Lampson, C. P., Tytell, A. A., Field, A. K., Nemes, M. M. & Hilleman, M. R. (1967). Inducers of interferon and host resistance. I. Double stranded RNA from extracts of Penicillum funiculosum. Proceedings of the National Academy of Science, U.S.A. 58, 782–9.CrossRefGoogle Scholar
Layne, E. (1957). Protein estimation with the Folin—Ciocalteau reagent. Methods in Enzymology 3, 448450.Google Scholar
Martin, R. C. & Ames, B. N. (1961). A method for determining the sedimentation behaviour of enzymes: application to protein mixtures. Journal of Biological Chemistry 236, 137 2–9.CrossRefGoogle ScholarPubMed
Mejbaum, W. (1939). Über die Bestimmung kleiner Pentosemengen, ins besondere in Derivaten der Adenylsaüre. Hoppe Seyler‘s Zeitschrift für Physiologische Chemie 258, 117–20.CrossRefGoogle Scholar
Merritt, K. & Johnson, A. C. (1965). Studies on the adjuvant action of bacterial endotoxins on antibody formation. VI. Enhancement of antibody formation by nucleic acids. Journal of Immunology 94, 4 1622.Google Scholar
Miles, A. A. & Pirie, N. W. (1939 a). The properties of antigenic preparations from Brucella melitensis. I. Chemical and physical properties of bacterial fractions. British Journal of Experimental Pathology 20, 8398.Google Scholar
Miles, A. A. & Pirie, N. W. (1939 b). The properties of antigenic preparations from Brucella melitensis. II. Serological properties of the antigens. British Journal of Experimental Pathology 20, 109–21.Google Scholar
Miles, A. A. & Pirie, N. W. (1939 c). The properties of antigenic preparations from Brucella melitensis. III. The biological properties of the antigen and the products of gentle hydro lysis. British Journal of Experimental Pathology 20, 278–96.Google Scholar
Morris, J. A. (1973). The use of polyacrylamide gel electrophoresis in taxonomy of brucella. Journal of General Microbiology 76, 23 1–7.Google Scholar
Osborn, M. J. (1963). Studies on the Gram-negative cell wall. I. Evidence for the role of 2-keto-3-deoxyoctonate in the lipopolysaccharide of Salmonella typhi-murium. Proceedings of the National Academy of Sciences, U.S.A. 50, 499506.Google Scholar
Paterson, J. S., Pirie, N. W. & Starbleforth, A. W. (1947). Protective antigens isolated from Br. abortus. British Journal of Experimental Pathology 28, 223–36.Google ScholarPubMed
Phillips, J. H., Braun, W. & Plescia, O. J. (1958). Antigenicity of a bacterial deoxyribonucleic acid. Nature, London 181, 573–5.CrossRefGoogle ScholarPubMed
Plescia, O. J., Noval, J., Palczuk, N. C. & Braun, W. (1961). Fractionation and irnmunological properties of a DNA-rich preparation from Brucella abortus. Proceedings of the Society for Experimental Biology and Medicine 106, 748–52.Google Scholar
Ralston, D. J. & Elberg, S. S. (1961). Intramonocytic destruction of brucella: potentiating effect of glycine on intracellular lysozyme activity. Journal of Infectious Diseases 109, 7180.Google Scholar
Renoux, G. (1964). Étalonnage des antigènes pour le diagnostic biologique de Ia brucellose. In Progress in Immunobiotogical Standardization 1, 176–84.Google Scholar
Scheidegger, J. J. (1955). Une microméthode de l‘immuno-électrophorèse. International Archives of Allergy and Applied Immunology 7, 103–10.Google Scholar
Smith, H., Keppie, J., Pearce, J. H. & Witt, K. (1962). The chemical basis of the virulence of Brucdlla abortus. IV. Immunogenic products from Brucella abortus grown in vivo. British Journal of Experimental Pathology 43, 538–48.Google ScholarPubMed
Snyder, F. & Stephens, N. (1959). A simple spectrophotometric determination of ester groups in lipids. Biochimica et Biophysica Acta 34, 244–7.Google Scholar
Tal, M. & Elson, D. (1963). The location of ribonuclease in Escherichia coli. Biochimica et Biophysica Acta 76, 4047.Google Scholar